RESUMO
Oenothein B (OeB), a dimeric ellagitannin with a macrocyclic structure, is reported to have beneficial effects, including antioxidant, antitumor, antiviral, and antimutagenic effects, on human health. Despite the remarkable properties of OeB, its role in neovascularization process has not yet been evaluated. Thus, this study aimed to evaluate the angiogenic activity of OeB using a chorioallantoic membrane (CAM) assay at different concentrations (6.25, 12.5, and 25 µg/µL), employing digital imaging and histological analysis. Furthermore, to elucidate the mechanisms by which OeB influences angiogenesis, we assessed the levels of vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNF-α) in CAM using immunohistochemical analysis. All concentrations of OeB significantly increased (p < 0.05) the percentage of vascularization as well as the levels of all the angiogenesis-associated parameters evaluated, indicating the pronounced pro-angiogenic activity of OeB. Our results showed that inflammation was one of the most relevant phenomena observed in CAM histology along with angiogenesis. In addition, a significant increase in VEGF and TNF-α levels was observed in all the CAMs compared to the negative control (p < 0.05). We suggest that OeB may induce the presence of inflammatory cells in CAM, leading to increased VEGF and TNF-α levels that result in the induction of angiogenesis. Therefore, OeB presents a favorable profile that could be further explored for the development of drugs for pro-angiogenic and tissue repair therapies.
Assuntos
Membrana Corioalantoide , Taninos Hidrolisáveis , Folhas de Planta , Fator de Necrose Tumoral alfa , Fator A de Crescimento do Endotélio Vascular , Fator de Necrose Tumoral alfa/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Animais , Folhas de Planta/química , Membrana Corioalantoide/efeitos dos fármacos , Taninos Hidrolisáveis/farmacologia , Embrião de Galinha , Eugenia/química , Indutores da Angiogênese/farmacologia , Neovascularização Fisiológica/efeitos dos fármacosRESUMO
Dioclea violacea seed mannose-binding lectin (DvL) has attracted considerable attention because of its interesting biological activities, including antitumor, antioxidant, and anti-inflammatory activities. This study evaluated the cytotoxic effect of DvL on tumor and normal cells using the mitochondrial activity reduction (MTT) assay, the carcinogenic and anti-carcinogenic activity by the epithelial tumor test (ETT) in Drosophila melanogaster, and the anti-angiogenic effect by the chick embryo chorioallantoic membrane (CAM) assay. Data demonstrated that DvL promoted strong selective cytotoxicity against tumor cell lines, especially A549 and S180 cells, whereas normal cell lines were weakly affected. Furthermore, DvL did not promote carcinogenesis in D. melanogaster at any concentration tested, but modulated DXR-induced carcinogenesis at the highest concentrations tested. In the CAM and immunohistochemical assays, DvL inhibited sarcoma 180-induced angiogenesis and promoted the reduction of VEGF and TGF-ß levels at all concentrations tested. Therefore, our results demonstrated that DvL is a potent anticancer, anti-angiogenic, and selective cytotoxic agent for tumor cells, suggesting its potential application as a prototype molecule for the development of new drugs with chemoprotective and/or antitumor effects.
Assuntos
Dioclea , Drosophila melanogaster , Neovascularização Patológica , Animais , Neovascularização Patológica/tratamento farmacológico , Neovascularização Patológica/metabolismo , Humanos , Dioclea/química , Embrião de Galinha , Drosophila melanogaster/efeitos dos fármacos , Carcinogênese/efeitos dos fármacos , Inibidores da Angiogênese/farmacologia , Membrana Corioalantoide/efeitos dos fármacos , Membrana Corioalantoide/irrigação sanguínea , Lectinas de Plantas/farmacologia , Células A549 , Linhagem Celular Tumoral , Camundongos , AngiogêneseRESUMO
Pain is a phenomenon present in the majority of the population, affecting, among others, the elderly, overweight people, and especially recently operated patients, analgesia being necessary. In the specific case of relief of postoperative pain, different kinds of anesthetics are being used, among them bupivacaine, a widely used drug which promotes long-lasting analgesic effects. However, cardiotoxicity and neurotoxicity are related to its repetitive use. To overcome these shortcomings, Novabupi® (a racemic mixture) was developed and is marketed as an injectable solution. This formulation contains an enantiomeric excess of the levogyre isomer, which has reduced toxicity effects. Seeking to rationalize its use by extending the duration of effect and reducing the number of applications, the objectives of this work were to develop and evaluate liposomes containing Novabupi (LBPV), followed by incorporation into thermogel. Liposomes were prepared using the lipid hydration method, followed by size reduction using sonication, and the developed formulations were characterized by hydrodynamic diameter, polydispersity index (PDI), surface zeta potential, and encapsulation efficiency. The selected optimal liposomal formulation was successfully incorporated into a thermogel without loss of thermoresponsive properties, being suitable for administration as a subcutaneous injection. In the ex vivo permeation studies with fresh rodent skin, the thermogel with liposomes loaded with 0.5% LBPV (T-gel formulation 3) showed higher permeation rates compared to the starting formulation, thermogel with 0.5% LBPV (T-Gel 1), which will probably translate into better therapeutic benefits for treatment of postoperative analgesia, especially with regard to the number of doses applied.
Assuntos
Analgesia/métodos , Levobupivacaína/administração & dosagem , Levobupivacaína/farmacocinética , Dor/tratamento farmacológico , Dor/metabolismo , Animais , Bovinos , Galinhas , Membrana Corioalantoide/efeitos dos fármacos , Membrana Corioalantoide/metabolismo , Géis , Humanos , Lipossomos , Masculino , Camundongos , Células NIH 3T3 , Técnicas de Cultura de Órgãos , Ratos , Ratos Wistar , Pele/efeitos dos fármacos , Pele/metabolismo , Absorção Cutânea/efeitos dos fármacos , Absorção Cutânea/fisiologiaRESUMO
Historically, the ocular toxicity of manufactured consumer materials has been evaluated using the rabbit in vivo Draize rabbit eye test. The animal data obtained were used by the United Nations Globally Harmonized System of Classification and Labelling of Chemicals (UN GHS) to define the classification and labelling (C&L) for eye damage/irritation endpoint. However, the Draize test, a method which was never formally validated, has been widely criticized because of its technical limitations. In addition, ethical and economic issues and advances in scientific knowledge, and political and public pressures have made animal experimentation unsustainable. This scenario has consequently led to the development of nonanimal testing and protocols/approaches with considerable predictive value and relevance for humans. It is widely accepted that one single nonanimal method cannot cover all the criteria of damage/inflammation assessed by regulatory adopted in vivo animal testing. Thus, integrated testing strategies (ITS) have been proposed, including a tiered testing approach combining different nonanimal testing with different endpoints, which have been used for regulatory purposes, on a case-by-case basis and within integrated approaches to testing and assessment (IATA), to identify materials according to their ability to trigger eye damage. In particular, the top-down and bottom-up approaches have been recommended for the C&L of materials, which cause serious eye damage or eye irritation, respectively. This chapter describes detailed protocols for eye irritation testing based on cells (Short Time Exposure-STE, OECD No. 491/2017), a vascularized membrane (the Hen's Egg Test-Chorioallantoic Membrane-HET-CAM) and corneal tissue (Bovine Corneal Opacity and Permeability-BCOP, OECD No. 437/2017), which can be applied using top-down or bottom-up approaches. In addition, it suggests making a corneal histomorphometric evaluation as an additional parameter in the BCOP method to differentiate materials that cause serious eye tissue damage (UN GHS Cat. 1) from materials that have reversible eye irritation effects (UN GHS Cat. 2).
Assuntos
Alternativas aos Testes com Animais , Bioensaio , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/efeitos dos fármacos , Córnea/efeitos dos fármacos , Irritantes/toxicidade , Neuropatia Óptica Tóxica , Testes de Toxicidade , Animais , Bovinos , Linhagem Celular , Sobrevivência Celular/efeitos dos fármacos , Embrião de Galinha , Córnea/patologia , CoelhosRESUMO
BACKGROUND: Curcumin (CUR) is a bioactive compound with several proven pharmacological properties. However, the major limitation for therapeutic use of CUR is its low bioavailability. In this sense, an alternative to this question is the use of polymeric nanocapsules (NC) as drug/nutraceutical delivery systems. Thus, the aim of current study was to assess the effect of CUR-loaded NC and their different coatings in chick embryo model, evaluating angiogenic, teratogenic and oxidative stress parameters. METHODS: The physicochemical characterization of unloaded and loaded NC with different coatings: (U-NC (P80), U-NC (PEG), U-NC (EUD), U-NC (CS), CUR-NC (P80), CUR-NC (PEG), CUR-NC (EUD) and CUR-NC (CS)) were performed. After 9 days of incubation, eggs were treated (10 mL/kg eggs; via injection) with NC (unloaded and loaded with CUR) and CUR-solution. In sequence, hen's egg test-chorioallantoic membrane (HET-CAM), angiogenic assay, external abnormalities, weight of embryos and oxidative stress markers (TBARS, NPSH, ROS and CAT) were analyzed. RESULTS: CUR-NC (P80, PEG, EUD and CS) treatments caused antiangiogenic and non-teratogenic effects in chick embryo model. Still, CUR-NC (P80), CUR-NC (PEG), CUR-NC (EUD) and CUR-NC (CS) did not alter markers of oxidative stress (TBARS, NPSH, CAT) studied. Only CUR-NC (EUD) caused increase in ROS levels. CONCLUSION: Wherefore, these findings of present study represent a advance in research of drug/nutraceutical delivery systems.
Assuntos
Curcumina/farmacologia , Nanocápsulas , Estresse Oxidativo/efeitos dos fármacos , Polímeros/química , Inibidores da Angiogênese/administração & dosagem , Inibidores da Angiogênese/farmacologia , Inibidores da Angiogênese/toxicidade , Animais , Embrião de Galinha , Galinhas , Membrana Corioalantoide/efeitos dos fármacos , Curcumina/administração & dosagem , Curcumina/toxicidade , Sistemas de Liberação de Medicamentos , Ovos , Espécies Reativas de Oxigênio/metabolismoRESUMO
Purpose: Disturbances that affect the inside of the eyeball tend to be highly harmful since they compromise the homeostasis of this organ. Alongside this, the eyeball has several anatomical barriers that prevent the entry of substances. This way, diseases that affect the retina are among those that present greater difficulty in the treatment. In many cases, abnormal proliferation of blood vessels (neovascularization) occurs from the lower layers of the retina. This process damages its structure physiologically and anatomically, causing the rapid and irreversible loss of visual capacity. This work aims to develop nanosuspensions of quantum dots (QDs) conjugated to bevacizumab. Methods: Two types of QDs were produced by aqueous route, stabilized with chitosan conjugated to bevacizumab. The antiangiogenic activity was evaluated in the chorioallantoic membrane model, in which results indicated discrete activity at the doses tested. Samples were assessed for their biosafety in animals, after intravitreal administration, by means of electroretinography (ERG), intraocular pressure (IOP) measurement, histological, morphometric, and immunohistochemical evaluation. Results: No significant alterations were detected in ERG that suggests damage to retinal function by the samples. No significant changes in IOP were also detected. The histological sections did not show signs of acute inflammation, although there was evidence of late retinal damage. The immunohistochemical analysis did not detect any apoptotic bodies. Conclusion: Preliminary results suggest that QDs present potential applicability in ocular therapy, and it is necessary to better characterize their in vivo behavior and to optimize their dosage.
Assuntos
Inibidores da Angiogênese/farmacologia , Bevacizumab/farmacologia , Pontos Quânticos/uso terapêutico , Retina/patologia , Inibidores da Angiogênese/administração & dosagem , Inibidores da Angiogênese/uso terapêutico , Animais , Bevacizumab/administração & dosagem , Bevacizumab/uso terapêutico , Membrana Corioalantoide/efeitos dos fármacos , Contenção de Riscos Biológicos/normas , Eletrorretinografia/métodos , Imuno-Histoquímica/métodos , Pressão Intraocular/efeitos dos fármacos , Injeções Intravítreas , Masculino , Modelos Animais , Nanopartículas/química , Nanopartículas/uso terapêutico , Neovascularização Patológica/diagnóstico , Neovascularização Patológica/tratamento farmacológico , Pontos Quânticos/administração & dosagem , Pontos Quânticos/química , Ratos , Degeneração Retiniana/diagnóstico , Degeneração Retiniana/metabolismo , Suspensões/administração & dosagem , Suspensões/química , Suspensões/farmacocinética , Membro 15 da Superfamília de Ligantes de Fatores de Necrose Tumoral/farmacologia , Fator A de Crescimento do Endotélio Vascular/imunologiaRESUMO
The Hancornia speciosa latex reveals angiogenic, osteogenic, and anti-inflammatory properties, which present its potential for developing of wound healing drugs; however, the latex compounds responsible for angiogenesis remain unknown. One strategy to screen these active compounds is evaluation of latex fractions. This study aimed to obtain different fractions of latex and evaluate its angiogenic activity separately using the chick chorioallantoic membrane (CAM) assay. The serum (SE) fraction was responsible for angiogenesis, which was subject to biochemical characterization and computational simulations in order to understand the contribution of H. speciosa latex in wound healing process. Our results revealed weak antioxidant potential and absence of antimicrobial activity in the SE fraction. Phytochemical analysis identified chlorogenic acids (CGA) as the main compound of SE fraction. CGA bioactivity predictions identify different molecules associated with extracellular matrix (ECM) remodeling, such as metalloproteinases, which also are overexpressed in our CAM assay experiment. Docking simulations revealed the interactions between CGA and matrix metalloproteinase 2. In conclusion, SE latex fraction stimulates angiogenesis and may influence ECM remodeling. These properties may contribute to the wound healing process, and also confirm the widespread use of this plant.
Assuntos
Indutores da Angiogênese/farmacologia , Apocynaceae/química , Membrana Corioalantoide/efeitos dos fármacos , Matriz Extracelular/efeitos dos fármacos , Látex/farmacologia , Extratos Vegetais/farmacocinética , Indutores da Angiogênese/isolamento & purificação , Animais , Apocynaceae/classificação , Embrião de Galinha , Cromatografia Líquida de Alta Pressão , Látex/isolamento & purificaçãoRESUMO
PURPOSE: Bevacizumab (BCZ) is a recombinant monoclonal antibody that inhibits the biological activity of the vascular endothelial growth factor, which has an important role in angiogenesis for tumoral growth and progression. In this way, our objective was to develop chitosan-coated lipid-core nanocapsules functionalized with BCZ by an organometallic complex using gold-III. METHODS: The formulation was produced and characterized in relation to physicochemical characteristics. Furthermore, the antitumoral and antiangiogenic activities were evaluated against C6 glioma cell line and chicken embryo chorioallantoic membrane (CAM), respectively. RESULTS: Final formulation showed nanometric size, narrow polydispersity, positive zeta potential and gold clusters size lower than 2 nm. BCZ in aqueous solution (0.01-0.10 µmol L-1) did not show cytotoxic activity in vitro against C6 glioma cell line; although, MLNC-Au-BCZ showed cytotoxicity with a median inhibition concentration of 30 nmol L-1 of BCZ. Moreover, MLNC-Au-BCZ demonstrated cellular internalization dependent on incubation time and BCZ concentration. BCZ solution did not induce significant apoptosis as compared to MLNC-Au-BCZ within 24 h of treatment. CAM assay evidenced potent antiangiogenic activity for MLNC-Au-BCZ, representing a decrease of 5.6 times in BCZ dose comparing to BCZ solution. CONCLUSION: MLNC-Au-BCZ is a promising product for the treatment of solid tumors.
Assuntos
Inibidores da Angiogênese/química , Bevacizumab/química , Quitosana/química , Glioma/tratamento farmacológico , Ouro/química , Lipídeos/química , Nanocápsulas/química , Inibidores da Angiogênese/farmacologia , Animais , Apoptose/efeitos dos fármacos , Bevacizumab/metabolismo , Linhagem Celular Tumoral , Permeabilidade da Membrana Celular , Embrião de Galinha , Membrana Corioalantoide/efeitos dos fármacos , Complexos de Coordenação/química , Relação Dose-Resposta a Droga , Composição de Medicamentos/métodos , Hexoses/química , Humanos , Lectinas de Plantas/química , Polissorbatos/química , Proteínas de Soja/química , Propriedades de Superfície , Fator A de Crescimento do Endotélio Vascular/metabolismoRESUMO
Considering the successful employment of alternative methods for eye toxicity assessment of products for regulatory purposes, and the recent advances in Brazilian legislative scenario, which adopted the UN GHS classification system for agrochemical formulations toxicity assessment, there is an emerging demand for strategies that allow the evaluation of such products. Based on this, the present study aimed to address the applicability of a mechanistic-based defined approach for eye toxicity assessment of agrochemical formulations. It was investigated the opacity/permeability, depth and location of corneal injury in bovine cornea, and vascular events in chorioallantoic membrane induced for different Brazilian agrochemicals using a Sequential Testing Strategy (STS). Cytotoxicity induced by the agrochemical formulations was evaluated by Short Time exposure (STE) (OECD TG 491) assay (step 1), corneal injury was investigated by standard Bovine Corneal Opacity and Permeability (BCOP) (OECD TG 437) followed by histopathological evaluation (step 2), and Hen Chorionic-allantoic Membrane test (HET-CAM) was used to evaluate vascular injury (step 3). The results demonstrated that the proposed defined approach enabled a classification corresponding UN GHS classification of agrochemical formulations while minimizing the use of live animals. Therefore, this approach may be useful for categorization of agrochemicals in Brazil according to the new regulatory scenario.
Assuntos
Agroquímicos/toxicidade , Alternativas aos Testes com Animais , Membrana Corioalantoide/efeitos dos fármacos , Córnea/efeitos dos fármacos , Irritantes/toxicidade , Animais , Bioensaio , Brasil , Bovinos , Galinhas , Membrana Corioalantoide/irrigação sanguínea , Córnea/metabolismo , Córnea/patologia , Opacidade da Córnea/induzido quimicamente , Permeabilidade , Testes de Toxicidade Aguda/métodosRESUMO
BACKGROUND: Fungal infections are highly prevalent and are responsible for high rates of morbidity and mortality. In this context, the search for new treatment alternatives is very relevant. OBJECTIVES: Analyse chemical compounds for antifungal potential against dermatomycosis fungi. METHODS: The antifungal activity of 121 compounds, intermediates or derivatives of 1,3-bis(aryloxy)propane substituted at C-2 (111 compounds) and isothiouronium derivatives (10 compounds) was investigated through susceptibility tests, mechanism of action, toxicity and hydrogel incorporation. RESULTS: The compound 1,3-bis(3,4-dichlorophenoxy)propan-2-aminium chloride (2j) was the most active fungicide against dermatophytes and Candida spp., at very low concentrations (0.39-3.12 µg/mL), including action on resistant and multidrug-resistant clinical strains. Compound 2j has presented a promising toxicity profile, showing selectivity index >10, relative to human lymphocytes. The compound was classified as non-irritant by the HET-CAM test and did not cause histopathological alterations in pig ear skin, thus presenting an excellent perspective for topical application. 2j targets the fungal cell wall, which was confirmed by scanning electron microscopy, which also indicated the additional ability of 2j to inhibit the Candida albicans pseudohyphae formation and biofilm of Microsporum canis. Compound 2j was incorporated in a hydrogel with bioadhesive potential. The results of the human skin permeation showed that 2j remained significantly in the epidermis, ideally for the dermatomycosis treatment. CONCLUSIONS: Therefore, the compound 2j demonstrated the potential for antifungal drug development, with a action mechanism elucidated and already applied in a semisolid formulation as a new therapeutic option for fungal skin infections.
Assuntos
Antifúngicos/farmacologia , Arthrodermataceae/efeitos dos fármacos , Candida/efeitos dos fármacos , Linfócitos/efeitos dos fármacos , Propano/análogos & derivados , Animais , Antifúngicos/química , Sobrevivência Celular , Células Cultivadas , Galinhas , Membrana Corioalantoide/efeitos dos fármacos , Orelha Externa/efeitos dos fármacos , Epiderme/efeitos dos fármacos , Ergosterol/metabolismo , Feminino , Citometria de Fluxo , Humanos , Hidrogéis , Concentração de Íons de Hidrogênio , Concentração Inibidora 50 , Masculino , Testes de Sensibilidade Microbiana , Microscopia Eletrônica de Varredura , Propano/química , Propano/farmacologia , Reologia , Relação Estrutura-Atividade , SuínosRESUMO
Gliomas account for nearly 70% of the central nervous system tumors and present a median survival of approximately 12-17 months. Studies have shown that administration of novel natural antineoplastic agents is been highly effective for treating gliomas. This study was conducted to investigate the antitumor potential (in vitro and in vivo) of Miconia chamissois Naudin for treating glioblastomas. We investigated the cytotoxicity of the chloroform partition and its sub-fraction in glioblastoma cell lines (GAMG and U251MG) and one normal cell line of astrocytes. The fraction showed cytotoxicity and was selective for tumor cells. Characterization of this fraction revealed a single compound, Matteucinol, which was first identified in the species M. chamissois. Matteucinol promoted cell death via intrinsic apoptosis in the adult glioblastoma lines. In addition, Matteucinol significantly reduced the migration, invasion, and clonogenicity of the tumor cells. Notably, it also reduced tumor growth and angiogenesis in vivo. Moreover, this agent showed synergistic effects with temozolomide, a chemotherapeutic agent commonly used in clinical practice. Our study demonstrates that Matteucinol from M chamissois is a promising compound for the treatment of glioblastomas and may be used along with the existing chemotherapeutic agents for more effective treatment.
Assuntos
Antineoplásicos Fitogênicos/uso terapêutico , Cromonas/uso terapêutico , Glioblastoma/tratamento farmacológico , Animais , Antineoplásicos Fitogênicos/isolamento & purificação , Antineoplásicos Fitogênicos/farmacologia , Apoptose/efeitos dos fármacos , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Embrião de Galinha , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/efeitos dos fármacos , Cromonas/isolamento & purificação , Cromonas/farmacologia , Glioblastoma/irrigação sanguínea , Humanos , Melastomataceae , Potencial da Membrana Mitocondrial/efeitos dos fármacos , Neovascularização Patológica/tratamento farmacológico , Extratos Vegetais , Folhas de PlantaRESUMO
Some recent studies have shown that pirfenidone (PFD) has favorable results in the healing process of the cornea. However, PFD in solution exhibits short half-life after topical application, and in this context, a liquid crystal nanoparticle system containing PFD (PFD-LCNPs) was developed. The nanoparticles were characterized by transmission electron microscopy, atomic force microscopy, small angle X-ray diffraction and polarized light microscopy. The PFD-LCNPs had particle size and zeta potential of 247.3â¯nm and -33.60â¯mV (stores at 4⯰C), respectively, and 257.5â¯nm and -46.00â¯mV (stored at 25⯰C), respectively. The pH of the formulation was 6.9 and the encapsulation efficiency was 35.9%. The in vitro release profiles indicated that PFD sustained release from PFD-LCNPs for up to 12â¯h. In vitro study of ocular irritation (HET-CAM test) concluded that components of the formulation are well tolerated for ocular administration. Corneal re-epithelialization time after chemical burning was significantly reduced in rabbits treated with PFD-loaded LCNPs when compared to the group treated with a vehicle. In addition, the anti-inflammatory action of pirfenidone was observed by reducing myeloperoxidase activity (MPO) and inflammatory cells in the histology of the tissues of animals treated with PFD-LCNPs. These findings indicated that the PFD-LCNPs might have the potential for effective ocular drug delivery.
Assuntos
Analgésicos/administração & dosagem , Anti-Inflamatórios/administração & dosagem , Queimaduras Químicas/tratamento farmacológico , Queimaduras Oculares/tratamento farmacológico , Cristais Líquidos , Nanopartículas/administração & dosagem , Piridonas/administração & dosagem , Administração Oftálmica , Analgésicos/farmacocinética , Animais , Anti-Inflamatórios/farmacocinética , Queimaduras Químicas/metabolismo , Queimaduras Químicas/patologia , Embrião de Galinha , Membrana Corioalantoide/efeitos dos fármacos , Córnea/efeitos dos fármacos , Córnea/metabolismo , Córnea/patologia , Sistemas de Liberação de Medicamentos , Liberação Controlada de Fármacos , Estabilidade de Medicamentos , Queimaduras Oculares/induzido quimicamente , Queimaduras Oculares/metabolismo , Queimaduras Oculares/patologia , Feminino , Tamanho da Partícula , Peroxidase/metabolismo , Piridonas/farmacocinética , CoelhosRESUMO
As a new strategy for treatment of ductal carcinoma in situ, biocompatible and bioadhesive nanoemulsions for intraductal administration of the cytotoxic agent piplartine (piperlongumine) were optimized in this study. To confer bioadhesive properties, the nanoemulsion was modified with chitosan or hyaluronic acid. Tricaprylin was selected as the nanoemulsion non-polar phase due to its ability to dissolve larger drug amounts compared to isopropyl myristate and monocaprylin. Use of phosphatidylcholine as sole surfactant did not result in a homogeneous nanoemulsion, while its association with polysorbate 80 and glycerol (in a surfactant blend) led to the formation of nanoemulsions with droplet size of 76.5⯱â¯1.2â¯nm. Heating the aqueous phase to 50⯰C enabled sonication time reduction from 20 to 10â¯min. Inclusion of either chitosan or hyaluronic acid resulted in nanoemulsions with similar in vitro bioadhesive potential, and comparable ability to prolong mammary tissue retention (to 120â¯h) in vivo without causing undesirable histological alterations. Piplartine was stable in both nanoemulsions for 60â¯days; however, the size of loaded NE-HA was maintained at a similar range for longer periods of time, suggesting that this nanoemulsion may be a stronger candidate for intraductal delivery.
Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Dioxolanos/administração & dosagem , Glândulas Mamárias Animais/metabolismo , Nanopartículas/administração & dosagem , Piperidonas/administração & dosagem , Adesividade , Animais , Antineoplásicos Fitogênicos/química , Galinhas , Quitosana/administração & dosagem , Quitosana/química , Membrana Corioalantoide/efeitos dos fármacos , Dioxolanos/química , Vias de Administração de Medicamentos , Emulsões , Feminino , Glicerol/administração & dosagem , Glicerol/química , Ácido Hialurônico/administração & dosagem , Ácido Hialurônico/química , Nanopartículas/química , Fosfatidilcolinas/administração & dosagem , Fosfatidilcolinas/química , Piperidonas/química , Polissorbatos/administração & dosagem , Polissorbatos/química , Ratos Wistar , Pele/química , SuínosRESUMO
Bacterial keratitis is an ocular infection that can lead to severe visual disability. Staphylococcus aureus is a major pathogen of the eye. We recently demonstrated the strong antimicrobial activity of LyeTxI-b, a synthetic peptide derived from a Lycosa erithrognatha toxin. Herein, we evaluated a topical formulation (eye drops) containing LyeTxI-b to treat resistant bacterial keratitis. Keratitis was induced with intrastromal injection of 4 × 105 cells (4 µL) in New Zealand female white rabbits. Minimum inhibitory concentration (MIC) and biofilm viability were determined. LyeTxI-b ocular toxicity was evaluated through chorioallantoic membrane and Draize tests. One drop of the formulation (LyeTxI-b 28.9 µmol/L +0.5% CMC in 0.9% NaCl) was instilled into each eye four times a day, for a week. Slit-lamp biomicroscopy analysis, corneal histopathological studies and cellular infiltrate quantification through myeloperoxidase (MPO) and N-acetylglucosaminidase (NAG) detection were performed. LyeTxI-b was very effective in the treatment of keratitis, with no signs of ocular toxicity. Planktonic bacteria MIC was 3.6 µmol/L and LyeTxI-b treatment reduced biofilm viability in 90%. LyeTxI-b eliminated bacteria and reduced inflammatory cellular activity in the eyes. Healthy and treated animals showed similar NAG and MPO levels. LyeTxI-b is a potent new drug to treat resistant bacterial keratitis, showing effective antimicrobial and anti-inflammatory activity.
Assuntos
Antibacterianos/administração & dosagem , Peptídeos Catiônicos Antimicrobianos/química , Proteínas de Artrópodes/administração & dosagem , Infecções Oculares Bacterianas/tratamento farmacológico , Ceratite/tratamento farmacológico , Soluções Oftálmicas/administração & dosagem , Venenos de Aranha/administração & dosagem , Infecções Estafilocócicas/tratamento farmacológico , Administração Tópica , Animais , Antibacterianos/toxicidade , Proteínas de Artrópodes/toxicidade , Galinhas , Membrana Corioalantoide/efeitos dos fármacos , Olho/efeitos dos fármacos , Olho/imunologia , Olho/patologia , Feminino , Neutrófilos/efeitos dos fármacos , Neutrófilos/imunologia , Soluções Oftálmicas/toxicidade , Coelhos , Venenos de Aranha/toxicidade , Staphylococcus aureusRESUMO
Background/Aims: The selection of suitable raw materials in the cosmetic research and development is a key point, not only in order to obtain the expected results but also to avoid undesirable side effects. This study evaluated the in vitro toxicity potential of four different plant extracts and their in vivo acceptability studies. Methods: Spirulina, Palmaria palmata, Cichorium intybus and Medicago sativa extracts were analysed alone or in combination and added in cosmetic formulations. The in vitro toxicity evaluation, Hen's Egg Chorioallantoic Membrane Test (HET-CAM) and 3T3 NRU phototoxicity test were performed to evaluate in vitro potential ocular irritation and photo safety, respectively. Twenty subjects were enrolled in the acceptability studies, who were evaluated for the absence of harmful effects of the formulation by visual assessment and by transepidermal water loss, a biophysical technique, for 30 days. Results: HET-CAM assay showed that the studied extracts added to a gel-cream formulation had no irritant potential. In addition, the combination of Palmaria palmata, alfalfa and chicory extracts did not show phototoxic potential in vitro. Acceptability studies showed that the formulation containing the four extracts combined did not provoke any transepidermal water loss (TEWL) alteration, sensory irritation or erythema in the forearms for the period of analysis. Conclusion: The studied active ingredients, alone or in combination, present no cytotoxicity potential and when added to a gel-cream formulation had no irritant potential in vitro. These results predicting no harmful effects were confirmed in the acceptability tests, which showed no alteration on skin barrier function and no report of irritation perception of sign of erythema, suggesting the potential of these extracts for the development of safe cosmetic products.
Assuntos
Cichorium intybus , Cosméticos/toxicidade , Medicago sativa , Extratos Vegetais/toxicidade , Rodófitas , Spirulina , Células 3T3 , Adulto , Animais , Galinhas , Membrana Corioalantoide/efeitos dos fármacos , Dermatite Fototóxica , Humanos , Camundongos , Pessoa de Meia-Idade , Pele/efeitos dos fármacos , Pele/metabolismo , Creme para a Pele , Testes de Toxicidade , Perda Insensível de Água/efeitos dos fármacosRESUMO
Growth differentiation factor 11 (GDF11) has been characterized as a key regulator of differentiation in cells that retain stemness features, despite some controversies in age-related studies. GDF11 has been poorly investigated in cancer, particularly in those with stemness capacity, such as hepatocellular carcinoma (HCC), one of the most aggressive cancers worldwide. Here, we focused on investigating the effects of GDF11 in liver cancer cells. GDF11 treatment significantly reduced proliferation, colony and spheroid formation in HCC cell lines. Consistently, down-regulation of CDK6, cyclin D1, cyclin A, and concomitant upregulation of p27 was observed after 24â¯h of treatment. Interestingly, cell viability was unchanged, but cell functionality was compromised. These effects were potentially induced by the expression of E-cadherin and occludin, as well as Snail and N-cadherin repression, in a time-dependent manner. Furthermore, GDF11 treatment for 72â¯h induced that cells were incapable of sustaining colony and sphere capacity in the absent of GDF11, up to 5â¯days, indicating that the effect of GDF11 on self-renewal capacity is not transient. Finally, in vivo invasion studies revealed a significant decrease in cell migration of hepatocellular carcinoma cells treated with GDF11 associated to a decreased proliferation judged by Ki67 staining. Data show that exogenous GDF11 displays tumor suppressor properties in HCC cells.
Assuntos
Proteínas Morfogenéticas Ósseas/farmacologia , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica , Fatores de Diferenciação de Crescimento/farmacologia , Neovascularização Patológica/prevenção & controle , Animais , Antígenos CD/genética , Antígenos CD/metabolismo , Proteínas Morfogenéticas Ósseas/genética , Proteínas Morfogenéticas Ósseas/metabolismo , Caderinas/genética , Caderinas/metabolismo , Diferenciação Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Embrião de Galinha , Membrana Corioalantoide/irrigação sanguínea , Membrana Corioalantoide/efeitos dos fármacos , Ciclina A/genética , Ciclina A/metabolismo , Ciclina D1/genética , Ciclina D1/metabolismo , Quinase 6 Dependente de Ciclina/genética , Quinase 6 Dependente de Ciclina/metabolismo , Inibidor de Quinase Dependente de Ciclina p27/genética , Inibidor de Quinase Dependente de Ciclina p27/metabolismo , Fatores de Diferenciação de Crescimento/genética , Fatores de Diferenciação de Crescimento/metabolismo , Células Hep G2 , Humanos , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologia , Ocludina/genética , Ocludina/metabolismo , Transdução de Sinais , Fatores de Transcrição da Família Snail/genética , Fatores de Transcrição da Família Snail/metabolismo , Esferoides Celulares/efeitos dos fármacos , Esferoides Celulares/metabolismo , Esferoides Celulares/patologiaRESUMO
Despite the standard approaches to treat the highly aggressive and invasive glioblastoma (GBM), it remains incurable. In this sense, cannabinoids highlight as a promising tool, because this tumor overexpresses CB1 and/or CB2 receptors and being, therefore, can be susceptible to cannabinoids treatment. Thus, this work investigated the action of the cannabinoid agonist WIN55-212-2 on GBM cell lines and non-malignant cell lines, in vitro and in vivo. WIN was selectively cytotoxic to GBM cells. These presented blebbing and nuclear alterations in addition to cell shrinkage and chromatin condensation. WIN also significantly inhibited the migration of GAMG and U251 cells. Finally, the data also showed that the antitumor effects of WIN are exerted, at least to some extent, by the expression of p53 and increased cathepsin D in addition to the decreased expression of HSP70.This data can indicate caspase-independent cell death mechanism. In addition, WIN decreased tumoral perimeter as well as caused a reduction the blood vessels in this area, without causing lysis, hemorrhage or blood clotting. So, the findings herein presented reinforce the usefulness of cannabinoids as a candidate for further evaluation in treatment in glioblastoma treatment.
Assuntos
Benzoxazinas/farmacologia , Glioblastoma/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Morfolinas/farmacologia , Naftalenos/farmacologia , Proteína Supressora de Tumor p53/metabolismo , Animais , Apoptose/efeitos dos fármacos , Caspases , Linhagem Celular , Movimento Celular/efeitos dos fármacos , Galinhas , Membrana Corioalantoide/efeitos dos fármacos , Membrana Corioalantoide/fisiologia , HumanosRESUMO
RESUMEN: El proceso angiogénico se define como el proceso en el que los vasos sanguíneos generan brotes dando como resultado neovascularidad. Un desbalance en el proceso angiogénico contribuye a numerosos desórdenes inflamatorios, infecciosos, isquémicos, inmunológicos y malignos. En el territorio maxilofacial se pueden encontrar patologías neoplásicas benignas de desarrollo local con un marcado componente angiogénico que determinan su crecimiento y agresividad. Sin embargo, existe escasa evidencia de cómo tratarlas en base al control de la angiogénesis. Terry & Jacoway (1994) desarrollaron un protocolo de tratamiento para lesiones neoplásicas benignas con un importante componente vascular que se utiliza actualmente. Este protocolo consiste en la infiltración intralesional de una suspensión de triamcinolona 10 mg/ml más una solución de anestésico local de uso odontológico como la lidocaína al 2 % asociada a epinefrina en una concentración de 1:200.000. Sin embargo, el uso de epinefrina podría disminuir la acción antiangiogénica de la triamcinolona al ser un vasoconstrictor. El objetivo de este estudio es comparar el efecto antiangiogénico, en la membrana alantocoriónica de pollo (MAC), de esta suspensión versus el efecto de la triamcinolona sin asociar a anestésicos locales. Los resultados del efecto antiangiogénico en la MAC de pollo, obtenidos en la investigación concluyeron que la suspensión de triamcinolona asociada a lidocaína con epinefrina es similar al de la suspensión de triamcinolona sin asociar a anestésicos locales. Además, se logró determinar que las suspensiones de triamcinolona sin asociar a anestésicos locales y las asociadas a anestésicos locales con o sin vasoconstrictor poseen un marcado efecto antiangiogénico, en la MAC de pollo, en comparación al grupo control.
SUMMARY: Angiogenesis is defined as the process through which new blood vessels form from previously existing vessels. Several inflammatory, infectious, ischemic, immunological and malignant disorders are caused by the lack of adequate angiogenesis balance. In the maxillofacial area, there are invasive benign neoplastic pathologies with a strong angiogenic component, which determines aggressive behavior and growth. Studies in the literature are scarce regarding treatment of these conditions based on angiogenesis control. Currently, the protocol used to treat these maxillofacial benign neoplastic lesions, was developed in 1994 by Terry & Jacoway and has a strong angiogenic component. Consequently lesions are treated via intra-lesion administration of triamcinolone 10 mg / mL, a solution used in dental local anesthetic, such as lidocaine 2 %, in conjunction with epinephrine at a concentration of 1:200,000. The objective of this study was to compare the antiangiogenic effect of this protocol in chicken chorioallantoic membrane (CAM) without the use of local anesthetic. The results of the antiangiogenic effect in the CAM obtained in this study concluded that the effect of the suspension of triamcinolone associated to lidocaine with epinephrine, is similar to the suspension of triamcinolone without associating local anesthetics. Furthermore, it was determined that suspensions of triamcinolone without local anesthetic, and those associated to local anesthetic with, and without vasoconstrictor have a strong antiangiogenic effect in CAM compared to the control group.
Assuntos
Animais , Embrião de Galinha , Triancinolona/administração & dosagem , Epinefrina/administração & dosagem , Inibidores da Angiogênese/administração & dosagem , Membrana Corioalantoide/efeitos dos fármacos , Lidocaína/administração & dosagem , Anestésicos Locais/administração & dosagem , Neovascularização PatológicaRESUMO
OBJECTIVES: The aim of this study was to evaluate the antifungal, antichemotactic and antioxidant activities of Schinus lentiscifolius essential oil, as well as its combined effect with terbinafine and ciclopirox, against dermatophytes. METHODS: Essential oil was analysed by GC-MS. The antifungal activity and the mechanism of action were determined by broth microdilution, sorbitol and ergosterol assays, as well as scanning electron microscopy. The checkerboard method was used for evaluating the interactions with commercial antifungal agents. The antioxidant and antichemotactic activities were measured using the DPPH and the modified Boyden chamber methods, respectively. KEY FINDINGS: Chemical analysis revealed the presence of 33 compounds, the primary ones being γ-eudesmol (12.8%) and elemol (10.5%). The oil exhibited 97.4% of antichemotactic activity and 37.9% of antioxidant activity. Antifungal screening showed effect against dermatophytes with minimum inhibitory concentration values of 125 and 250 µg/ml. Regarding the mechanisms of action, the assays showed that the oil can act on the fungal cell wall and membrane. Synergistic interactions were observed using the combination with antifungals, primarily terbinafine. CONCLUSIONS: Schinus lentiscifolius essential oil acted as a chemosensitizer of the fungal cell to the drug, resulting in an improvement in the antifungal effect. Therefore, this combination can be considered as an alternative for the topical treatment of dermatophytosis.